The manufacture of clinical grade gene transfer vectors requires the development of efficient, large scale methods that are appropriate for the generation of safe materials to be used in human study subjects. This work involves both laboratory expertise and regulatory oversight of the safety of the products made in the Human Gene Therapy Applications Laboratory (HGTAL). Specific Aim 1: The HGTAL will work collaboratively with the AAV preclinical vector development core, Directed by Dr. Xiao Xiao, to translate laboratory AAV production techniques into a scalable and controlled manufacturing process that can be used to produce clinical grade AAV in years 2 and 3 to support the proposed clinical trial that is projected to start in year 3. Specific Aim 2: The HGTAL will manufacture and certify a lot of recombinant AAV for the support of a phase 1 dose determination study. This material will also be certified and characterized according to CBER guidelines and will be used supporting toxicology studies. Specific Aim 3: The HGTAL will refine and improve upon its AAV vector manufacturing capabilities and will produce at least two subsequent lots of recombinant AAV vector to support a phase II efficacy study. A determination of the size of the lots needed to be produced will be finalized based upon the highest dose which can be administered to patients, as determined in the phase 1 dose escalation study.